Journal Article

AAV-HBV mouse model replicates the intrahepatic immune landscape of chronic HBV patients at single-cell level

Kukolj, George, Beyens, Matthias, Krishna, Vinod, Berge, Koen Van den, Van Gulck, Ellen, Conceição-Neto, Nádia, Han, Qinglin, Podlaha, Ondřej, Pierson, Wim, Zhu, Ren, Dockx, Koen, Nájera, Isabel, Wu, Qun, De Maeyer, Dries, Aerts, Liese, Li, Chris, Yao, Zhiyuan

2025

Overview

Unresolved hepatitis B virus (HBV) infection leads to a progressive state of HBV-specific immune dysfunctionality that characterizes chronic infection. The immune-competent adeno associated virus (AAV)-HBV mouse model is commonly used preclinically, though a comprehensive characterization of the liver immune microenvironment and its translatability to human infection is still lacking. We investigated the intrahepatic immune profile of the AAV-HBV mouse model at a single-cell level and compared with data from CHB patients in immune tolerant (IT) and immune active (IA) clinical stages. Immune exhaustion was profiled through an iterative subclustering approach for cell-typing analyses of single-cell RNA-sequencing data in CHB donors and compared to the AAV-HBV mouse model 4-weeks and 24-weeks post-transduction to assess its translatability. This was confirmed using an exhaustion flow cytometry panel at 4 and 42-weeks post-transduction. Using single-cell RNA-sequencing, CD8 pre-exhausted T-cells with self-renewing capacity ( ), and terminally exhausted CD8 T-cells ( ) were detected in the AAV-HBV model. These terminally exhausted CD8 T-cells (expressing ) were significantly enriched versus control mice and independently identified through flow cytometry. Importantly, comparison to CHB human data showed a similar exhausted CD8 T-cell population in IT and IA donors, but not in uninfected individuals. Long term high titer AAV-HBV mouse liver transduction led to T-cell exhaustion, as evidenced by expression of conventional immune checkpoint markers at mRNA and protein levels. In both IT and IA donors, a similar CD8 exhausted T-cell population was identified, with increased frequency observed in IA donors. These data support the use of the AAV-HBV mouse model to study classical T-cell exhaustion in HBV infection and the effect of immune-based therapeutic interventions.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: AAV-HBV-infected mouse model / Animals / Antigens / CD8 antigen / CD8-Positive T-Lymphocytes - immunology / chronic HBV / Chronic infection / Dependovirus - genetics / Dependovirus - immunology / Disease Models, Animal / Ethics / Female / Flow cytometry / Hepatitis B / hepatitis B virus / Hepatitis B virus - genetics / Hepatitis B virus - immunology / Hepatitis B, Chronic - immunology / Hepatitis B, Chronic - virology / Humans / Immune checkpoint / Immunology / Liver / Liver - immunology / Liver - virology / Lymphocytes / Lymphocytes T / Male / Mice / Microenvironments / mRNA / Single-Cell Analysis / single-cell transcriptome / T cell exhaustion / Therapeutic applications / Transduction / Veins & arteries / Viral infections / Viruses